This proposal focuses on the development of multi-well ELISA-like assays to quantify DNA binding activity of two breast cancer prognosis markers, Sp1 and ER, with much more sensitivity and high throughput than standard DNA gel shift techniques. Such an approach has already been successfully applied to monitoring transcription factor activation by Active Motif in the TransAM(TM) product line. However, none of these kits has been validated for diagnostic purpose. We will use a DNA probe immobilized onto a 96-well microplate to capture Sp1 or ER protein in an extract from cells or tissues. An antibody specific for Sp1 or ER will identify the bound protein and yield a quantitative results in two hours or so. Such assay will overcome the disadvantages of gel shift and allow many cancer and age research laboratories to establish the clinical importance of Sp 1 and ER function. The ultimate aim of this proposal is the development of a new assay that provides a fast and effective test for determining and predicting whether the hormonal treatment is the appropriate therapy for the particular cancer. PROPOSED COMMERCIAL APPLICATION: Early and accurate prognosis of cancer is key to patient survival. For breast, uterine, ovary and prostate cancer patients, the efficiency of hormonal therapy is based on the presence of estrogen and progesterone receptors (ER/PR) in the tumor. This proposal focuses on the development of mltiwell ELISA-like assays to quantify DNA binding activity of two breast cancer prognosis markers, Sp1 and ER, with much more sensitivity and high throughput than standard DNA gel shift techniques. Making such technology available for Sp1 and ER proteins for research and clinical facilities would speed up breast turmorigenesis knowledge. Developing the assay as a fast and effective test for predicting whether the hormonal treatment is the apppropriate therapy for the patient would target 500,000 new cancer cases a year.